{"product_id":"espcas9-bhn20152546","title":"eSpCas9","description":"\u003ch2\u003eOverview\u003c\/h2\u003e\n\u003cp\u003eThis GenCefe mRNA encodes \u003cstrong\u003eeSpCas9\u003c\/strong\u003e, a cas protein construct supplied for gene editing and genome engineering applications. The product is formulated as lyophilised, non-encapsulated RNA and is intended for use in cell-based research applications requiring transient protein expression with reduced immunogenicity.\u003c\/p\u003e\n\u003ch2\u003emRNA Construct Design\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003e\n\u003cstrong\u003e5′ Cap:\u003c\/strong\u003e Cap1 (m7GpppNm) — co-transcriptionally added during in vitro transcription (IVT). Cap1 includes 2′-O-methylation at the first transcribed nucleotide, closely mimicking the cap structure found on endogenous mammalian mRNA and reducing recognition by innate immune sensors (e.g., IFIT1\/IFIT3).\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eModified Nucleotides:\u003c\/strong\u003e 100% N1-methylpseudouridine (m1Ψ; N1-Me-Pseudo UTP) substitution for all uridine residues. m1Ψ modification reduces TLR7\/TLR8-mediated innate immune activation and PKR-driven translational suppression, resulting in improved protein expression in immunocompetent cells and primary cell types.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003ePoly(A) Tail:\u003c\/strong\u003e 100–120 nt — enzymatically polyadenylated. The poly(A) tail stabilises the 3′ terminus, supports poly(A)-binding protein (PABP) recruitment, and enhances ribosome recycling for efficient cap-dependent translation.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003e5′ UTR:\u003c\/strong\u003e hHBA1 (hemoglobin subunit alpha 1 5′ UTR) — a well-characterised human UTR that supports efficient cap-dependent translation initiation.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003e3′ UTR:\u003c\/strong\u003e hHBA1 (hemoglobin subunit alpha 1 3′ UTR) — provides post-transcriptional stability and modulates mRNA decay kinetics.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eSignal Peptide:\u003c\/strong\u003e No\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eProtein Tag:\u003c\/strong\u003e No\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eCodon Optimisation:\u003c\/strong\u003e No (native human codon usage retained)\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003emRNA Length:\u003c\/strong\u003e Provided upon order placement.\u003c\/li\u003e\n  \u003cli\u003e\n\u003cstrong\u003eForm:\u003c\/strong\u003e Lyophilised powder; reconstitute in DEPC-treated water as needed.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003cp\u003eThis mRNA is supplied as non-encapsulated, lyophilised powder. Delivery vehicle selection (LNP, electroporation, lipofection) is at the discretion of the end user and should be optimised for the target cell type and application.\u003c\/p\u003e\n\u003ch2\u003eBiological Background\u003c\/h2\u003e\n\u003cp\u003eCRISPR-associated (Cas) proteins are programmable RNA-guided endonucleases derived from bacterial adaptive immune systems. Class 2 Cas nucleases, including the widely used \u003cem\u003eStreptococcus pyogenes\u003c\/em\u003e Cas9 (SpCas9) and the Cpf1 (Cas12a) orthologues from \u003cem\u003eAcidaminococcus\u003c\/em\u003e (AsCas12a\/AsCpf1) and \u003cem\u003eLachnospiraceae\u003c\/em\u003e (LbCas12a\/LbCpf1), introduce site-specific double-strand breaks guided by single guide RNA (sgRNA) or CRISPR RNA (crRNA). Delivery of Cas nuclease as mRNA rather than plasmid DNA reduces the risk of off-target genomic integration, restricts editing activity to the transient window of mRNA half-life, and minimises innate immune responses in primary cells, making mRNA-based CRISPR delivery increasingly preferred for therapeutic genome editing and ex vivo haematopoietic stem cell (HSC) modification.\u003c\/p\u003e\n\u003ch2\u003eResearch Relevance and Current Trends\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003eNon-integrating genome editing: Cas9 and Cas12a mRNA co-delivery with guide RNA (as synthetic crRNA:tracrRNA duplexes or sgRNA) achieves transient editing activity, reducing the off-target editing window compared to plasmid-based systems.\u003c\/li\u003e\n  \u003cli\u003eBase editing and prime editing: Modified Cas protein mRNAs (e.g., high-fidelity SpCas9 variants) serve as the nuclease backbone for base editors (CBEs\/ABEs) and prime editors, where mRNA delivery reduces immunogenicity and enables editing in post-mitotic cells.\u003c\/li\u003e\n  \u003cli\u003eClinical-grade HSC editing: Ex vivo haematopoietic stem cell editing programmes (e.g., for haemoglobinopathies) favour mRNA-RNP delivery for regulatory safety profiles in GMP-adjacent workflows.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003ch2\u003eCommon Research Applications\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003eTransient genome editing — Cas9\/Cas12a mRNA co-delivery with guide RNA for site-directed insertions, deletions, or substitutions in primary cells and cell lines.\u003c\/li\u003e\n  \u003cli\u003eIndel analysis — mRNA-based editing followed by surveyor assay, Sanger sequencing, or NGS to quantify on-target editing efficiency at defined genomic loci.\u003c\/li\u003e\n  \u003cli\u003eBase editing and prime editing — Cas mRNA as the scaffold for base editor or prime editor fusion proteins for precise nucleotide conversions without double-strand breaks.\u003c\/li\u003e\n  \u003cli\u003eRNP optimisation — Cas protein mRNA-to-guide-RNA ratio titration to maximise editing efficiency while minimising cytotoxicity across cell types.\u003c\/li\u003e\n\u003c\/ul\u003e\n\u003ch2\u003eNotes for Experimental Interpretation\u003c\/h2\u003e\n\u003cul\u003e\n  \u003cli\u003eCas protein mRNA activity depends on guide RNA quality and design; evaluate multiple guide sequences across the target locus and confirm on-target editing by sequencing before committing to functional experiments.\u003c\/li\u003e\n  \u003cli\u003eHigh Cas9 mRNA doses can be cytotoxic, particularly in primary cells; titrate mRNA amount empirically and assess viability 24 h post-transfection before proceeding to editing analysis.\u003c\/li\u003e\n  \u003cli\u003eeSpCas9 (enhanced specificity variant) contains specific mutations (e.g., K848A\/K1003A\/R1060A) that reduce off-target activity; confirm the variant designation matches the intended specificity profile.\u003c\/li\u003e\n  \u003cli\u003eCas12a (Cpf1) orthologs have distinct PAM requirements (TTTV for LbCas12a; TTTV\/TTTT for AsCas12a); verify PAM compatibility at the target site before designing crRNA.\u003c\/li\u003e\n\u003c\/ul\u003e","brand":"GenCefe Biotech","offers":[{"title":"20 ug","offer_id":53239633084781,"sku":"IR0101002","price":279.0,"currency_code":"USD","in_stock":true},{"title":"100 ug","offer_id":53239639998829,"sku":"IR0101010","price":339.0,"currency_code":"USD","in_stock":true},{"title":"200 ug","offer_id":53239640031597,"sku":"IR0101020","price":399.0,"currency_code":"USD","in_stock":true},{"title":"500 ug","offer_id":53239640064365,"sku":"IR0101050","price":459.0,"currency_code":"USD","in_stock":true},{"title":"1 mg","offer_id":53239640097133,"sku":"IR0101100","price":549.0,"currency_code":"USD","in_stock":true},{"title":"5 mg","offer_id":53239640129901,"sku":"IR0101500","price":1799.0,"currency_code":"USD","in_stock":true}],"thumbnail_url":"\/\/cdn.shopify.com\/s\/files\/1\/0949\/7424\/7277\/files\/mRNA_fae889b1-5a2d-4f68-a5d6-858524792794.png?v=1776677729","url":"https:\/\/www.ebiohippo.com\/products\/espcas9-bhn20152546","provider":"BioHippo","version":"1.0","type":"link"}